活性氧及能量代谢障碍在体外培养肝细胞脂肪变性中的作用

Role of ROS and energy metabolism dysfunction in hepatocyte adipose degeneration in vitro

目的:探讨细胞活性氧(ROS)及线粒体能量代谢障碍在乙醇和小牛血清诱导的体外培养肝细胞脂肪变性中的作用。方法:将人肝细胞株L02进行细胞培养,加入不同浓度的乙醇,采用MTT法确定乙醇作用的最佳浓度;用2%乙醇和50%小牛血清(A+CS)诱导L02细胞,以油红O染色观察细胞内脂滴形成状况,并检测细胞TG的含量;用流式细胞仪(FCM)测定细胞活性氧及线粒体膜电位的改变;采用反相高效液相色谱分析法测定细胞ATP含量。结果:2%乙醇和50%小牛血清作用36h,肝细胞内有典型的脂滴形成,与对照组相比,细胞内TG含量及ROS水平明显增加;而细胞线粒体膜电位及肝细胞ATP的含量则显著降低(P<0.01)。结论:在乙醇和小牛血清诱导肝细胞脂肪变过程中,细胞ROS的增加及线粒体能量代谢障碍可能发挥了重要的作用。

AIM： To explore the role of reactive oxygen species （ROS） and energy metabolism dysfunction in hepatocyte adipose degeneration induced by alcohol and calf serum （CS）. METHODS： The growing L02 cells were treated with different concentrations of alcohol. To screen the proper concentration of alcohol, the proliferation of cells was measured by MTF. Lipid droplets in the cells were observed through oil red staining. Triglyceride （TG） content was detected with analyzed kit. The level of ROS and changes of mitochondrial membrane potential （Δψm） in cells were tested by flow cytometry. Reversed - phase high performance liquid chromatography （ RP - HPLC） was applied to assay the cellular ATP content. RESULTS： Lipid droplets were observed under light microscope in the cells treated with 2% alcohol and 50% CS（A ＋ CS） for 36 h. Compared to control group, the cellular TG and ROS levels in model group markedly increased while Δψm and ATP content in cells significantly decreased （P 〈 0. 01 ）. CONCLUSION： ROS and energy metabolism dysfunction may play an important role in hepatocyte adipose degeneration induced by alcohol and CS.